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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1997

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
no
Principles of method if other than guideline:
Adminstration of test material
1 or 2 times per gavage 24 hours apart
1 x by intraperitoneal route
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Carbon tetrachloride
- Physical state: liquid
- Lot/batch No.: purchased from Wako Pure Chemical Co, Osaka, Japan

Test animals

Species:
mouse
Strain:
other: BDF1
Sex:
male

Administration / exposure

Route of administration:
other: 1 or 2 x per gavage, alternatively 1 x ip
Duration of treatment / exposure:
one or two gavage treatments 24 hours apart for bone marrow examination
single ip treatment for peripheral blood assay
Frequency of treatment:
one or two gavage treatments 24 hours apart for bone marrow examination
single ip treatment for peripheral blood assay
Post exposure period:
24 hours after last treatment for bone marrow assessment
24, 48 and 72 hours after ip injection for peripheral blood assay
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
1000, 2000 and 3000 mg/kg intraperitoneal
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
500, 1000 and 2000 mg/kg by gavage
Basis:
nominal conc.
No. of animals per sex per dose:
5 per dose and treatment schedule
Control animals:
yes, concurrent vehicle

Examinations

Tissues and cell types examined:
Bone marrow cell smears from femur, slides fixed with methanol and stained with 2.5 % Giemsa solution.
5 microL of blood placed on acridine orange-coated slide and covered.

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Negative controls validity:
not specified
Positive controls validity:
valid

Any other information on results incl. tables

Bone marrow test (Table 1):

There was no increase in the frequency of micronucleated polychromatic erythrocytes due to the treatment of carbon tetrachloride after either one or two treatments. Mitomycin C (single treatment) produced a statistically significant increase at all three time points .

Table 1: Bone marrow micronucleus test of carbon tetrachloride with oral gavage:

 Dose (mg/kg) No of mice No of injections  mean Micronuc. PCEs   PCE ratios   
0 (corn oil)  0.20 +/- 0.16  38.1 +/- 5.3  
500  0.20 +/- 0.10  27.4 +/- 6.2 
1000  0.38 +/- 0.18  29.7 +/- 10.7 
2000  0.22 +/- 0.16  24.8 +/- 5.0 
         
Mitomycin C 0.5 mg/kg  1.58 +/- 0.61 (p< 0.01)  39.7 +/- 3.2 
         
0 (corn oil)   0.12 +/- 0.04  36.0 +/- 3.5 
500  0.28 +/- 0.16  26.0 +/- 3.9 
1000  0.28 +/- 0.22  22.5 +/- 9.6 
2000  0.20 +/- 0.19  20.8 +/- 8.6 

Peripheral blood test (Table 2):

There was no inclrease in the frequency of micronucleated reticulcytes due to the teatment of carbon tetrachloride at any observation time point. Mitomycin C produced a statistically significant increase

Table 2: Peripheral blood micronucleus test of carbon tetrachloride with intraperitoneal injection

 Dose (mg/kg) No of mice  Treatment time  % Micronucl. RET, means +/- SD  
1000  0.20 +/- 0.07 
  24  0.24 +/- 0.09 
  48  0.20 +/- 0.12 
  72  0.26 +/- 0.17 
       
2000  0.18 +/- 0.13 
  24  0.18 +/- 0.16 
  48  0.26 +/- 0.11 
  72  0.28 +/- 0.13 
       
3000  0.16 +/- 0.09 
  24  0.26 +/- 0.05 
  48  0.20 +/- 0.10 
  72  0.20 +/- 0.16 
       
Mitomycin C  0.16 +/- 0.09 
1 mg/kg  24  0.56 +/- 0.34 (p<0.01) 
  48  3.90 +/- 0.67 (p<0.01) 
  72  0.96 +/- 0.37 (p<0.01) 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
Carbon tetrachloride does not induce micronuclei and thus devoid of a mutagenic potential in this test system.
Executive summary:

Carbon tetrachloride was investigated in a mouse micronucleus test. Administrations were once or twice (24 hours apart) by gavage and sampling 24 hours later for the bone marrow assay or once by the ip route with samplings after 0, 24, 48 and 72 hours for the peripheral blood assay.

In none of the investigations were the micronucleated polychromatic erythrocytes (bone marrow) or micronucleated reticulocytes (peripheral blood) increased at any time point, while Mitmycin C showed the expected increases.

Carbon tetrachloride is devoid of a mutagenic potential in this test system.